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Regulation of transferrin-induced indocytosis by wildetype and C282Y-mutant HFE in transfected HeLa cells

MPG-Autoren
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Henkel,  Andreas Wolfram
Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society;

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Both,  Matthias
Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society;

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Migala,  Andrea
Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society;

Externe Ressourcen

http://ajpcell.physiology.org/content/282/5/C973.full.pdf+html
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http://dx.doi.org/10.1152/ajpcell.00415.2001
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Zitation

Schwake, L., Henkel, A. W., Riedel, H. D., Schlenker, T., Both, M., Migala, A., et al. (2002). Regulation of transferrin-induced indocytosis by wildetype and C282Y-mutant HFE in transfected HeLa cells. American Journal of Physiology: Cell Physiology, 282, 973-979. doi:10.1152/ajpcell.00415.2001.


Zitierlink: https://hdl.handle.net/11858/00-001M-0000-0029-221A-6
Zusammenfassung
The hereditary hemochromatosis protein HFE is known to complex with the transferrin receptor; however, its function regarding endocytosis of transferrin is unclear. We performed patch-clamp capacitance measurements in transfected HeLa cells carrying wild-type or C282Y-mutant HFE cDNA under the control of a tetracycline-sensitive promoter. Whole cell experiments in cells with suppressed expression of wild-type HFE revealed a decrease in membrane capacitance, reflecting predominance of endocytosis in the presence of transferrin. Cells overexpressing C282Y-mutant HFE displayed less intense capacitance decreases, whereas no significant decrease was observed in cells overexpressing wild-type HFE. The formation of single endocytic vesicles in cells with suppressed expression of wild-type HFE was greatly increased in the presence of transferrin as revealed by cell-attached recordings. According to their calculated diameters, many of these vesicles corresponded to clathrin-coated vesicles. These results suggest that wild-type HFE negatively modulates the endocytic uptake of transferrin. This inhibitory effect is attenuated in cells expressing C282Y-mutant HFE. Time-resolved measurements of cell membrane capacitance provide a powerful tool to study transferrin-induced endocytosis in single cells.